Brazilian Archives of Biology and Technology

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An overview on molecular markers for detection of ochratoxigenic fungi in coffee beans
(Instituto de Tecnologia do Paraná - Tecpar, 2009-11) Fungaro, Maria Helena Pelegrinelli; Sartori, Daniele
Ochratoxin A (OTA) is one of the most abundant food-contaminating mycotoxins. Its occurrence in several agricultural commodities has been considered a worldwide problem. This toxin is mainly produced by Aspergillus species. OTA has nephrotoxic, immunosuppressive, and carcinogenic effects and consequently the contamination with this toxin represents a high risk for human and animal health. In the last 5 years, several investigators have applied molecular methods in order to develop PCR assays for identifying and quantifying OTA-producing fungi in coffee beans samples. The main objective is to allow the detection of microorganisms capable of producing OTA, preferentially prior to ochratoxin production and accumulation. In this contribution several of these attempts will be reviewed and discussed.
Caffeine Formation by Suspension Cultures of Coffea dewevrei
(Instituto de Tecnologia do Paraná - Tecpar, 2000-01) Sartor, Rosana Mary; Mazzafera, Paulo
The low caffeine content in leaves of C. dewevrei (~ 0.5 mg/g) is due to a low biosynthesis associated with a fast degradation. On the other hand, high biosynthesis and low degradation confer a higher content (~ 8 mg/g) in leaves of C. arabica. In this work it was observed that cell cultures of C. dewevrei recovered the ability to synthesize caffeine almost in similar levels of C. arabica cultures. Tracer experiments with labelled carbon dioxide showed a significant accumulation of radioactivity in caffeine and metabolites, indicating an active biosynthesis. When the cultures were fed with labelled caffeine most of the radioactivity was recovered in caffeine, indicating that although active, degradation was not so efficient as in leaves, and therefore, contributing for the alkaloid accumulation in the cell cultures.
Characterization of Roasted Coffea arabica Species by the Relationship Between Caffeine and Diterpenes Contents
(Instituto de Tecnologia do Paraná - Tecpar, 2020) Zanin, Rodolfo Campos; Kitzberger, Cíntia Sorane Good; Benassi, Marta de Toledo
Commercial roasted and ground coffees are usually blends of Coffea arabica and Coffea canephora. Considering the differences in price and sensory characteristics between these two species, the identification of the presence of each species in commercial blends is of great interest. The aim of this study was to describe typical profiles of caffeine and diterpenes (kahweol and cafestol) contents and the ratios among these compounds to support the characterization of Coffea species in roasted coffees. 32 good cup quality Brazilian C. arabica coffees (from coffee quality contests) produced using different postharvest treatments were studied. All analysis were performed by HPLC. Higher ranges were observed in diterpene contents – kahweol varied from 1.75 to 10.68 g/kg (coefficient of variation of 510%) and cafestol from 1.76 to 9.66 g/kg (449%) – than caffeine, that varied from 5.1 to 16.2 g/kg (coefficient of variation of 218%). Wide ranges of the kahweol/cafestol ratio (0.63 to 2.77) and the caffeine/kahweol ratio (0.84 to 5.15) were also observed. Hence it was proposed the additional use of a new parameter, the ratio of caffeine/sum of diterpenes (kahweol + cafestol) that presents values from 0.54 to 2.39. The results indicated that the combined use of these parameters could be a potential tool for discriminating Coffea species in blends of roasted and ground coffee. It was proposed as potentially indicative of C. arabica: values of kahweol/cafestol ratio above 0.50, associated with caffeine/kahweol ratio lower than 5.50 and caffeine/sum of diterpenes ratio lower than 2.50.
Characterization of the biomass of a hybrid anaerobic reactor (HAR) with two types of support material during the treatment of the coffee wastewater
(Instituto de Tecnologia do Paraná - Tecpar, 2013-05) Silva, Vivian Galdino da; Campos, Cláudio Milton Montenegro; Pereira, Erlon Lopes; Silva, Júlia Ferreira da
This study investigated the microbiology of a hybrid anaerobic reactor (HAR) in the removal of pollutant loads. This reactor had the same physical structure of an UASB reactor, however with minifilters inside containing two types of support material: expanded clay and gravel. Two hydraulic retention times (HRT) of 24h and 18h were evaluated at steady-state conditions, resulting in organic loading rates (OLR) of 0.032 and 0.018 kgDBO 5 m -3 d -1 and biological organic loading rates (BOLR) of 0,0015 and 0.001 kgDBO 5 kgSVT -1 d -1 , respectively. The decrease in concentration of organic matter in the influent resulted an endogenous state of the biomass in the reactor. The expanded clay was the best support material for biofilm attachment.
Coffee berry borer resistance in coffee genotypes
(Instituto de Tecnologia do Paraná - Tecpar, 2010-03) Sera, Gustavo Hiroshi; Sera, Tumoru; Ito, Dhalton Shiguer; Ribeiro Filho, Claudionor; Villacorta, Amador; Kanayama, Fabio Seidi; Alegre, Clayton Ribeiro; Grossi, Leandro Del
The aim of this study was to evaluate the coffee germplasm of the Paraná Agronomic Institute (IAPAR) for resistance to the coffee-berry-borer. Preliminary field evaluation was performed in August 2004 and the fruits of less damaged genotypes in the field were evaluated under controlled condition with obligated and free choice experiments established in a randomized complete design with three replications. The genotypes were evaluated fifteen days after infestation with one borer per fruit in Petri dishes. The data were analyzed by the Scott-Knott means test at 1 % and by the χ 2 test. Statistical analysis indicated that Coffea kapakata, Psilanthus bengalensis, C. eugenioides and genotypes with C. eugenioides genes were resistant. These genotypes presented low frequency of bored grains. C. eugenioides and C. kapakata could present resistance at epicarp level but not in the grain. P. bengalensis could present resistance also in the grains.
A comparison between enzyme immunoassay and HPLC for ochratoxin A detection in green, roasted and instant coffee
(Instituto de Tecnologia do Paraná - Tecpar, 2007-03) Fujii, Simone; Ono, Elisabete Yurie Sataque; Ribeiro, Ricardo Marcelo Reche; Assunção, Fernanda Garcia Algarte; Takabayashi, Cássia Reika; Oliveira, Tereza Cristina Rocha Moreira de; Itano, Eiko Nakagawa; Ueno, Yoshio; Kawamura, Osamu; Hirooka, Elisa Yoko
An indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) for ochratoxin A (OTA) detection in green, roasted and instant coffees was developed using anti-OTA monoclonal antibody. Immunological reagents prepared were OTA-BSA (4.76 μ g/mL), anti-OTA.7 MAb (2x10 3 -fold dilution) and HRP-anti IgG (10 3 -fold dilution). The detection limit was 3.73 ng OTA/g and correlation coefficients (r) between this immunoassay and high performance liquid chromatography were 0.98 for green coffee, 0.98 for roasted and 0.86 for instant. OTA levels detected by ic- ELISA were higher than by HPLC, with ELISA/HPLC ratio of 0.66 - 1.46 (green coffee), 0.96 - 1.11 (roasted) and 0.93 - 1.82 (instant). ELISA recoveries for OTA added to coffee (5 - 70 ng/g) were 81.53 % for green coffee, 46.73 % for roasted and 64.35 % for instant, while recoveries by HPLC were 80.54 %, 45.91 % and 55.15 %, respectively. Matrices interferences were minimized by samples dilution before carrying out the ELISA assay. The results indicate that MAb-based ic-ELISA could be a simple, sensitive and specific screening tool for OTA detection, contributing to quality and safety of coffee products.
Comparison of high performance liquid chromatography with fluorescence detector and with tandem mass spectrometry methods for detection and quantification of ochratoxin a in green and roasted coffee beans
(Instituto de Tecnologia do Paraná - Tecpar, 2013-11) Bandeira, Raquel Duarte da Costa Cunha; Uekane, Thais Matsue; Cunha, Carolina Passos da; Rodrigues, Janaina Marques; Cruz, Marcus Henrique Campino de la; Godoy, Ronoel Luiz de Oliveira; Fioravante, Andreia de Lima
Two analytical methods for the determination and confirmation of ochratoxin A (OTA) in green and roasted coffee samples were compared. Sample extraction and clean-up were based on liquid–liquid phase extraction and immunoaffinity column. The detection of OTA was carried out with the high performance liquid chromatography (HPLC) combined either with fluorescence detection (FLD), or positive electrospray ionization (ESI+) coupled with tandem mass spectrometry (MS–MS). The results obtained with the LC-ESI-MS/MS were specific and more sensitive, with the advantages in terms of unambiguous analyte identification, when compared with the HPLC-FLD.
Compositional changes of proteins and amino acids in germinating coffee seeds
(Instituto de Tecnologia do Paraná - Tecpar, 2000-07) Shimizu, Milton Massao; Mazzafera, Paulo
Endosperm is the main reserve tissue in coffee seeds. Coffee (Coffea arabica L.) seeds were germinated for six weeks and qualitative and quantitative changes in amino acids and proteins were investigated. The total content of free amino acids were reduced during germination, however, protein content remained constant. SDS-PAGE profiles showed that legumin-like proteins became less stained in the last weeks. Asparagine, glutamic acid, aspartic acid, alanine and lysine were the major free amino acids, although serine and glutamine were also significant. Except for tyrosine, which increased with germination, all other amino acids were reduced. Analysis of the amino acid composition of the total soluble protein showed glutamic acid/glutamine and glycine as the main amino acids. However, other amino acids such as leucine, aspartic acid/asparagine, alanine, lysine, serine were also found in reasonable amounts.
Development of Adsorbent Materials Using Residue from Coffee Industry and Application in Food dye Adsorption Processes
(Instituto de Tecnologia do Paraná - Tecpar, 2022-11-11) Castro, Luiz Eduardo Nochi de; Battocchio, Débora Alessandra Jones; Ribeiro, Leomara Floriano; Colpini, Leda Maria Saragiotto
The aim of this study was to develop an adsorbent from coffee husk, residue from the coffee industry by different activation methods and to evaluate its application in the treatment of effluent containing synthetic Ponceau 4R dye. The activations were with phosphoric acid and zinc chloride, then the samples were characterized in terms of physical and chemical composition, structure and morphology, thermal behavior and kinetic and thermodynamic study. Regarding the characterization of the material, the activation with phosphoric acid showed porous morphology and a high surface area (429.7 m2 g-1) and with the kinetic study it was possible to obtain an equilibrium time of 60 min and dispersion according to Elovich kinetics (R2 = 0.96 and RMSE < 0.24). According to the experimental design, the adsorptive capacity can be evaluated with the significant result (qe = 19.87 mg g-1) under the conditions of 90 min of the adsorption process and at the concentration of 2 g L-1 of adsorbent. In relation to the equilibrium study the Langmuir model fit the data with coefficient of determination of R2 = 0.84. Finally, the Ponceau 4R dye adsorption process using coffee husks as an activated carbon was thermodynamically favorable through the negative Gibbs free energy and positive entropy (+0.022 kJ mol-1 K-1). Therefore, the use of coffee husk as activated carbon proved to be a favorable material for Ponceau 4R dye adsorption.
Dietary utilisation of protein and energy from fresh and ensiled coffee pulp by the nile tilapia, oreochromis niloticus
(Instituto de Tecnologia do Paraná - Tecpar, 2003-03) Moreau, Yann; Arredondo, Jose-Luis; Gaime, Isabelle Perraud; Roussos, Sevastianos
Dietary protein and energy utilisation of diets containing fresh and ensiled coffee pulp were studied on 3.2 ± 0.2 g Nile tilapia for 28 days. Diets formulation and feeding were designed on the basis of daily dietary protein and energy allowance. A control diet A (100 % protein and 100 % energy allowance) corresponding to 15 g CP kg -1 day -1 and 750 kJ kg -1 day -1 , a low protein control diet B (80 % protein and 100 % energy allowance), two diets C and E (100 % protein and 100 % energy allowance) where 20 % of protein were supplied by coffee pulp, and two diets D and F with the same amount of coffee pulp than in C and E and supplementation in non-protein energy. Inclusion of coffee pulp in the diet strongly impaired growth and feed utilisation. Silage process improved overall feed utilisation comparing to fresh coffee pulp. Results showed that fresh or ensiled coffee pulp was not a suitable feedstuff for Nile tilapia. However, better knowledge on modification occurring during silage process could allow finding the way to significantly improve nutritive value of coffee pulp by-products.
Dynamic Behavior of Coffee Branches: an Analysis Using the Finite Element Method
(Instituto de Tecnologia do Paraná - Tecpar, 2024-01-12) Melo Junior, Wellington Washington Andrade de; Santos, Fábio Lúcio; Scinocca, Francisco; Rosa, Pablo Antunes da; Magalhães, Ricardo Rodrigues
The use of computational simulation techniques is an important tool for the coffee harvesting issues, particularly the finite element method. The method is widely used in the structural analysis of agricultural machinery, as well as in the analysis of the stresses and vibrations of coffee branches and peduncles during the harvesting process. The present study aimed to develop three-dimensional finite element models of the plagiotropic branches of the Catuaí Vermelho variety of Arabica coffee in different positions along the orthotropic branches of the plant; considering high-fidelity models. Additionally, by considering the branches’ experimental properties (physical-mechanical), the natural frequencies and vibration modes of the branches were determined by means of computer simulations. First, the geometric properties of the coffee branches were obtained by means of two images taken using a professional camera to obtain the input data of the virtual simulation. For the mechanical properties, it was used a semi-analytical digital scale, to obtain the mass of the specimens. The modulus of elasticity was determined using a universal testing machine. The variability in the simulated natural frequencies could be identified, which was on the order of 30% for the first frequency, regardless of the position of the branch in the plant. These values were lower for the other frequencies. Linear regression fits showed a coefficient of determination, and correlation tests were used to verify the relationship between the values obtained numerically and experimentally, which were validated by using experimental data using the modal analysis techniques.
Effect of leaf water potential on cold tolerance of Coffea arabica L.
(Instituto de Tecnologia do Paraná - Tecpar, 2002-12) Caramori, Lázara Pereira Campos; Caramori, Paulo Henrique; Manetti Filho, João
Young coffee plants from cultivar Mundo Novo of Coffea arabica were grown without irrigation for 32 consecutive days, to evaluate the effect of leaf water potential on damage caused by low temperatures, under controlled conditions. À wide range of leaf water potentials were evaluated, from — 0.45MPa (wet soil) at the beginning of the experimental period, to — 4.8MPa (severe leaf wilting) at the end. Results showed that under moderate water stress, there was a higher frequency of undamaged plants and lower frequency of severely damaged plants. These results help explain part of the regional variability observed after a frost and stress the importance of new studies associating cold and drought tolerance in coffee.
Enzyme Production by Induratia spp. Isolated from Coffee Plants in Brazil
(Instituto de Tecnologia do Paraná - Tecpar, 2020) Monteiro, Mônica Cristina Pereira; Tavares, Dérica Gonçalves; Nery, Eduardo Mateus; Queiroz, Marisa Vieira de; Pereira, Olinto Liparini; Cardoso, Patrícia Gomes
Endophytic fungi belonging to the genus Muscodor now transferred to Induratia are known producers of bioactive volatile organic compounds (VOCs) with many industrial applications. However, the members of this genus have rarely been reported to produce non-volatile metabolites including enzyme. Enzymes of the endophytes are degraders of the polysaccharides available in the host plants and the knowledge of enzyme production by Induratia spp. may provide insights into their possible biotechnological applications. The aim of this study was to evaluate the activity of amylase, cellulase, lipase, pectinase, phytase, protease, endo β-1,4 glucanase and exo β-1,4 glucanase enzymes produced by fungi of the species Induratia coffeana, Induratia yucatanensis and Induratia sp. isolated from organic coffee plants. All Induratia spp. were able to produce the extracellular enzymes cellulase, pectinase, protease, and phytase. Eight fungi were able to produce lipase and four produced amylase. The specific activity of endo β-1, 4 glucanase and exo β-1,4 glucanase enzymes were detected for 9 and 8 endophytic fungi, respectively. This work demonstrated for the first time, the array of enzymes produced by Induratia spp. isolated from Coffea arabica in organic systems in Brazil.
Evaluation of arsenic and selenium in brazilian soluble coffee by inductively coupled plasma atomic emission spectrometry with hydride generation
(Instituto de Tecnologia do Paraná - Tecpar, 2001-10) Santos, Éder José dos; Oliveira, Elisabeth de
A method for the evaluation of arsenic and selenium in soluble coffee by inductively coupled plasma atomic emission spectrometry with continuous hydride generation to attend the Brazilian food legislation is described. Samples were digested with nitric acid and hydrogen peroxide in a focused microwave system. Slow heating eliminated nitric acid and selenium (VI) was reduced to selenium (IV) by addition of 6 mol/L hydrochloric acid and heating at 90°C under a reflux system. The influence of sample acidity on sensitivity was investigated. Hydrochloric acid 6 mol/L was the most suitable reaction medium. Practical detection limits of 2.0μg/L for As and 1.0μg/L for Se were achieved and attended the Brazilian food legislation. The results of recoveries on spiked samples demonstrate the reliability and accuracy of the procedure.
Genetic diversity among forty coffee varieties assessed by rapd markers associated with restriction digestion
(Instituto de Tecnologia do Paraná - Tecpar, 2005-07) Diniz, Leandro Eugênio Cardamoni; Ruas, Claudete de Fátima; Carvalho, Valdemar de Paula; Torres, Fabrício Medeiros; Ruas, Eduardo Augusto; Santos, Melissa de Oliveira; Sera, Tumoru; Ruas, Paulo Maurício
The genetic variability of 40 accessions of C. arabica was evaluated using a combination of the random amplified polymorphic DNA (RAPD) technique and restriction digestion of genomic DNA. The genetic variability and the relatedness among all accessions were initially evaluated using 195 RAPD primers which revealed a very low level of genetic variation. To improve the efficiency in the detection of polymorphism, the genomic DNA of all accessions were submitted to digestion with restriction endonucleases prior to PCR amplification. A total of 24 primers combined with restriction digestion of DNA rendered 318 bands, of which 266 (83.65%) were polymorphic. The associations among genotypes were estimated using UPGMA-clustering analysis. The accessions were properly clustered according to pedigree and agronomic features. The ability to distinguish among coffee accessions was greater for RAPD plus restriction digestion than for RAPD alone, providing evidences that the combination of the techniques was very efficient for the estimation of genetic relationship among C. arabica genotypes.
Genetic variability for frost resistance among coffea accessions assessed in the field and in a cold chamber
(Instituto de Tecnologia do Paraná - Tecpar, 2005-01) Petek, Marcos Rafael; Sera, Tumoru; Alteia, Marcos Zorzenon
The genetic variability for frost resistance was evaluated in field conditions and cold chamber among Coffea accessions. Results showed that C. liberica var. dewevrei and C. racemosa, as well as hybrids obtained from these species were more resistant to frost. There was a great genetic variability for frost resistance among the evaluated sources (b 2 = 0,98). The correlation estimated between damages in the field and in the cold chamber was 0.933**, therefore, this methodology was efficient and feasible and could be potentially used in a breeding program.
High metal contents in coffee plant organs developed in tubets with different proportions of biosolid composts and carbonized rice hulls
(Instituto de Tecnologia do Paraná - Tecpar, 2004-08) Franco, Elias; Miglioranza, Edison; Fonseca, Ésio de Pádua
The objective of this study was to analyze the effect of biosolid composts (BC) and carbonized rice hull (RH) on the production of Coffea arabica L. cv Catuaí Vermelho seedlings (IAC H 2077-2-5-99) grown in tubets. A totally randomized experimental design with five treatments and four replications was conducted in a nursery , to evaluate Co, Ni, Cr, Pb, Cu, Mn and Zn contents in roots, stems and leaves. Higher contents of Cr, Co, Ni, Pb, Cu and Zn were found in roots, while leaves carried the highest content of Mn. Zn in leaves went from average to high in plants developed in BC. In plants developed in RH. Mn content was about four times higher than the adequate concentration. Ni, Co, Cr, Pb and Cu contents found in leaves were considered normal, according to international standards. The best treatment for coffee seedling nutrition was 50% RH + 50% BC.
Influence of coffee pruning on the severity of frost damage
(Instituto de Tecnologia do Paraná - Tecpar, 2000-03) Androcioli Filho, Armando; Caramori, Paulo Henrique
Frost damages in a field experiment of pruning types and systems for the cultivars of Coffea arabica Catuaí and Mundo Novo, were evaluated at Londrina (23 o 22’S, 52 o 10 ́W), State of Parana, southern Brazil, during the winter of 1990 and 1994. Pruning types evaluated were ‘esqueletamento’ (cutting off all plagiotropic branches at 20-30 cm from the orthotropic branch), ‘decote’ (cutting off the orthotropic branch at 1.5 m and 2.0 m above ground) and ‘recepa’ (cutting off the orthotropic branch at 0.8 m above ground), performed on all rows and on alternate rows, and on different sections of the plant. Results indicated that frost damage could increase according to the type and height of pruning. The pruning type ‘esqueletamento’ and prunings at higher levels were more suitable for regions with frost risk. Under severe frost condition, pruning type did not affect the damage in anyone of the treatments evaluated.
Influence of integral and decaffeinated coffee brews on metabolic parameters of rats fed with hiperlipidemic diets
(Instituto de Tecnologia do Paraná - Tecpar, 2013-09) Gomes, Júlia Ariana de Souza; Faria, Bruno Generoso; Silva, Viviam de Oliveira; Zangerônimo, Márcio Gilberto; Miranda, José Rafael; Lima, Adriene Ribeiro de; Pereira, Rosemary Gualberto Fonseca Alvarenga; Pereira, Luciano José
The objective of this study was to evaluate the influence of integral and decaffeinated coffee brews (Coffea arabica L and C. canephora Pierre) on the metabolic parameters of rats fed with hyperlipidemic diet. Thirty male Wistar rats (initial weight of 270 g ± 20 g) were used in the study, which were divided into six groups five each. The treatments were normal diet, hyperlipidemic diet, hyperlipidemic diet associated with integral coffee arabica or canephora brews (7.2 mL/kg/day) and hyperlipidemic diet associated to decaffeinated arabica, or canephora brews, using the same dosage. After 41 days, performance analyses were conducted.The rats were then euthanized and the carcasses were used for the analysis of dried ether extract and crude protein. Fractions of adipose tissue were processed for histological analysis. There was a reduction in weight gain and accumulation of lipids in the carcasses, lower diameter of adipocytes and a lower relative weight of the liver and kidneys of rats fed with hyperlipidemic diet associated with integral coffee brew. Integral coffee brew reduced the obesity in the rats receiving hyperlipidemic diet, but the same effect did not occur with the decaffeinated types.
Influence of Silver Nitrate on Somatic Embryogenesis Induction in Arabica Coffee (Coffea arabica L.)
(Instituto de Tecnologia do Paraná - Tecpar, 2019) Rojas-Lorz, Laura; Arrieta-Espinoza, Griselda; Valdez-Melara, Marta; Pereira, Luiz Filipe Protasio; Gatica-Arias, Andrés
The influence of silver nitrate (AgNO3), benzyladenine (BAP), and indole-3-acetic acid (IAA) on low frequency somatic embryogenesis (LFSE) induction in Caturra and Catuaí arabica coffee was evaluated. For the Caturra cultivar, the production of somatic embryos was significantly increased by adding AgNO3 to the semisolid culture medium. The highest average number of somatic embryos for this cultivar was obtained using 6.6 μM BAP, 2.85 μM IAA, and 40 μM AgNO3. In contrast, for the Catuaí cultivar, the highest average number of somatic embryos was obtained using semisolid medium supplemented with 8.8 μM BAP, and 2.85 μM IAA. Using these protocols, somatic embryos were directly induced using leaf sections of in vitro plants of both coffee cultivars within 8 weeks. The somatic embryos developed into rooted plants with a 100% survival rate upon transfer to the greenhouse.